Treated or untreated cancer cells were lysed in 0.5 ml of Tris-EDTA buffer, at pH 7.4, containing 0.2% (v/v) of triton x-100 and upon centrifugation for 10min at 13xg, 4°C, fragmented DNA will get separated from intact chromatin in a microfuge tube (labeled as B). Then the fragmented DNA, present in supernatant layer will be transferred to other tube (labeled as T). Then, 0.5 ml of 25% TCA will be filled into each B and T tube and vortexed vigorously. DNA gets precipitated upon keeping for overnight at 4°C and again centrifuge at 13× g at 4°C for 10 min. Supernatant will be discarded and add 80 μl of 5% TCA to each pellet.
5ml of the extract was pipetted into a 50ml flask and then 10ml of distilled water was added. 2ml of ammonium hydroxide solution and 5ml of concentrated amylachohol were made up to mark and left to react for 30min for colour development. This was measured at 505nm. Alkaloid determination using harborne (1973) method 5g of the sample was weighed into a 250ml beaker and 200ml of 10% acetic acid in ethanol was added and covered and allowed to stand for 4 hours. This was filtered and the extract was concentrated on a water bath to one quarter of the original volume.
The isolated rat's brain was fixed and processed as per the tissue processing procedure, and then brain tissue slices were prepared for toluidine blue staining. The slices were incubated with 0.5% (w/v) solution of toluidine blue for 20 minutes. After being dehydrated, cleared and mounted, brain slices were examined under a light microscope. Morphological changes in hippocampal neurons in CA1, CA3 and DG area were assessed microscopically by using 40x magnifications (Sadeghi et al,
After 30 minutes, tubes were taken out and kept in ice-cold water for 30 minutes. These were centrifuged at 3000 rpm for 15 minutes. The absorbance of the supernatant was read at 540 nm at room temperature against appropriate blank. Blank consist of 1 ml distilled water, 0.5 ml of 30% TCA, and 0.5 ml of 0.8% TBA. TBARS values were expressed as n moles malonaldehyde (MDA)/mg protein.
Incubated at 25℃ for 7 days under the shaking condition. After 7 days, centrifuge it at 5000rpm and at temperature of 4℃ for 20 minutes. Discard the pellet that contains bacterial cells, take supernatant. Make the pH of supernatant of 2 by using "M" "H" _2 "S" "O" _4. Add choloroform and methanol of ratio 2:1 and of equal volume.
1. Label each well of a tissue culture treated 6-well plate appropriately for each cell line or condition being investigated. 2. Prepare 2x cell culture medium by dissolving 1 g of powder medium and 0.2 g of sodium bicarbonate in de-ionized water to a final volume of 50 ml. 3.
Collection and Prepration of the Inoculum The fecal matter was collected from naturally infected sheep and it was examined for Eimeria oocysts by a coverglass flotation method using Sheather's sugar solution to concentrate the oocysts (Sloss and Kemp, 1978.). The infected fecal material was mixed in 2.5 % potassium dichromate solution and was left for sporulation at room temperature for one week. The sporulated oocysts were subsequently quantified using the Mc master technique (Maff, 1986). A strict morphological criterion was used to specify Eimeria species present depending on the morphometric character according to Levine and Ivens (1986). Experimental Animals Thirty healthy lambs aged from 4-6 months, free
bronchiseptica biofilm formation was evaluated by using a microtiter dish assay following the methodology described previously . For each strain, 100 µl of a bacterial suspension adjusted to an optical density at 650 nm (OD650) of 0.05 were inoculated into the corresponding wells of microtiter plate. Plates were incubated statically at 36oC for 24 or 48 h. For 48 h cultures, the growth medium was entirely replaced with a fresh one after 24 h. To quantify biofilm formation, the liquid medium containing planktonic bacteria was firstly removed from each well. The remaining adhered biomass in every well was gently washed twice with PBS and subsequently stained for 20 min with a 0.1 % p/v Cristal Violet (CV) solution. After staining the CV solution was removed and every well was washed twice with distilled water.
Imatinib and Nilotinib following tests are carried out; FISH (Fluorescence in situ hybridization), Bone marrow Aspirate (BMA), Polymerase chain reaction (PCR) and USG (Ultrasonography test) Study Method: A data acquisition form was filled about the demographic details, diagnosis, treatments and laboratory findings from patient medical records. CBC reports were assessed before initiating the drug therapy. Later, laboratory values were checked after each follow up. Each patient received an average dose of 400mg of Imatinib and 800mg of Nilotinib. This retrospective observational case study designed to interpret the effectiveness and toxicity of Imatinib and Nilotinib in CML patients.
2ml of 10% ammonium citrate was added to each beaker. The pH was then adjusted to 8.5 by adding 10 drops of 5M NH4OH(aq) to each beaker. 3ml of 0.1% cuprizone was added to each beaker. The four solutions were then transferred to 25ml volumetric flasks. The beakers were washed with de-ionized water, and the washings were combined with the solutions in the 25ml volumetric flask.